Anti-Poliovirus Receptor/PVR antibody [EPR17302] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-Poliovirus Receptor/PVR antibody [EPR17302] - BSA and Azide free (AB228348)

Poliovirus Receptor/PVR was immunoprecipitated from 1mg of U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate with ab205304 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab205304 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1 : U-87 MG whole cell lysate 10μg (Input).
Lane 2 : ab205304 IP in U-87 MG whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab205304 in U-87 MG whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 10 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205304).

All lanes:

Immunoprecipitation - Anti-Poliovirus Receptor/PVR antibody [EPR17302] (<a href='/en-us/products/primary-antibodies/poliovirus-receptor-pvr-antibody-epr17302-ab205304'>ab205304</a>)

Predicted band size: 45 kDa

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• WB

Lab

Western blot - Anti-Poliovirus Receptor/PVR antibody [EPR17302] - BSA and Azide free (AB228348)

This data was developed using the same antibody clone in a different buffer formulation (ab205304).

Lanes 1- 4 : Merged signal (red and green). Green - ab205304 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab205304 was shown to react with Poliovirus Receptor/PVR in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266102 (knockout cell lysate ab257622) was used. Wild-type HEK-293T and PVR knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab205304 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Poliovirus Receptor/PVR antibody [EPR17302] (<a href='/en-us/products/primary-antibodies/poliovirus-receptor-pvr-antibody-epr17302-ab205304'>ab205304</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PVR knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PVR (Poliovirus Receptor) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-pvr-poliovirus-receptor-knockout-hek-293t-cell-line-ab266102'>ab266102</a>)

Lane 3:

Wild-type A549 cell lysate at 20 µg

Lane 4:

PVR knockout A549 cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 70 kDa

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